scholarly journals PRODUKSI ANTIBODI POLIKLONAL HASIL INDUKSI ISOLAT PROTEIN NON KINASE DARI MEMBRAN SPERMATOZOA MANUSIA

1970 ◽  
Vol 15 (2) ◽  
Author(s):  
Umie Lestari ◽  
Basuki B Purnomo
Keyword(s):  
Molecular Weight ◽  
Protein Concentration ◽  
Tween 20 ◽  
Sds Page ◽  
Protein Membrane ◽  
Good Concentration ◽  
Elisa Technique ◽  
Biuret Method ◽  
Method Show

Objective: The aim of this research is to get the profile of human sperms membrane protein based on identification for molecular weight, concentration protein, phosphorylation activity, and ability to raise enough antibodies. Material and methods: There are three phases in this research, first, isolation protein of human sperms membrane with using lysis buffer containing Tween-20, second, determination of molecular weight, phosphorylation activity, and protein concentration, and third, antibody production. Molecular weight of protein membrane was detected by SDS-PAGE, confirmation of protein concentration measured Biuret method, kinase activity measured using spectrophometry at its optimum condition, and then production of antibody that corfirmed by ELISA technique. Results: Human sperms contain the protein with molecular weight of 201, 163, 116, 97, 72, 56, 48, 32, 27, 20, and 17kDa. The highest phosphorylation activity was found on protein with molecular weight of 20 kDa is 30.197x 10-3 unit, while the lowest phosphorylation activity is 28.976x10-3 unit was shown on protein with 116 kDa of molecular weight and then   this protein can induce production of antibody. Biuret method show the protein concentration from protein with molecular weight 116 kDa is 410 ug/mL. Conclusion: The protein that has molecular weight of 116 kDa can be used as immunocontraception agent because phosphorylation activity has lower, good concentration, and could induce the production of antibody.

scholarly journals PENGARUH POLISAKARIDA KRESTIN DARI EKSTRAK JAMUR Coriolus versicolor TERHADAP PROFIL PROTEIN TERSTIKULER DAN KADAR TESTOSTERON Mus musculus

el–Hayah ◽  
2016 ◽  
Vol 5 (4) ◽  
pp. 169
Author(s):  
Sri Puji Astuti Wahyuningsih ◽  
Virid Gibson ◽  
Alfiah Hayati
Keyword(s):  
Molecular Weight ◽  
Treatment Group ◽  
Mus Musculus ◽  
Control Group ◽  
Protein Profiles ◽  
Testosterone Levels ◽  
Randomized Design ◽  
Sds Page ◽  
Elisa Technique ◽  
Completely Randomized Design

<em>The purpose of this research was to determine the effect of polysaccharide krestin</em> (<em>PSK) </em><em>on the testicular protein profiles and testosterone levels of Mus musculus with variety of dosages. This research used a completely randomized design. It were devide into four treatment group i.e. control group, PSK treatment at a dose of  15, 30, 60 mg/kgBW. Each group had six replications. Testicular proteins were isolated by flushing technique and analized by SDS-PAGE. Testosterone levels were analized using ELISA technique at wavelength 450 nm. Protein bands analysis showed that there were no diversification between four treatments. Molecular weight of protein bands were 87, 63, 57, 35, and 30 kDa. The results of research showed that the testosterone levels at dosage 60 mg/kgBW had significanly different with control, PSK treatment of 15 and 30 mg/kgBW. PSK treatment of  60 mg/kgBW had lowest level at dosage, i.e. 25946.8 ρg/mL. It can be concluded that giving variety of dosages of polysaccharide krestin did not affect to testicular protein profiles but giving effect to testosterone levels of Mus musculus.</em>

Affinity Chromatography of Human Factor VIII Using Human and Rabbit Antibodies to Factor VIII

1979 ◽  
Vol 42 (04) ◽  
pp. 1306-1315 ◽  
Author(s):  
Janet L Lane ◽  
H Ekert ◽  
A Vafiadis
Keyword(s):  
Molecular Weight ◽  
Factor Viii ◽  
Affinity Chromatography ◽  
Protein Concentration ◽  
Gel Filtration ◽  
Subunit Structure ◽  
Molecular Weights ◽  
Human Factor Viii ◽  
Sds Page ◽  
Normal Human

SummaryFactor VIII, purified by gel filtration on Sepharose 2B, has an 8 band multiple subunit structure, with molecular weights ranging from 30,000 to 230,000, on reduction and SDS-PAGE at a protein concentration of 400 μg/gel. Affinity chromatography of this factor VIII preparation with insolubilized haemophilic antibody to factor VIII showed that 45-81% VIII:C and 0-33% VIILRag were attached to the column. Elution of the column with 0.25 M CaCl2 did not show VIII:C or VIILRag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed that 95 % of the protein bound by haemophilic antibody had a molecular weight similar to the low molecular weight subunits of the reduced factor VIII.In control experiments with normal Human IgG, 3% of VIII:C and 5% of VIILRag were attached to the column. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the protein, showed 2 faint bands with molecular weight consistent with heavy and light chains of IgG.Similar experiments with antibody to factor VIII showed that 67-83% of VIILC and 61-76% of VIII:Rag were attached to the column. Elution of the column with 0.25 M CaCl2 showed 10% of the applied VIII:C, but no VIII:Rag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed an 8 band subunit structure similar to the reduced factor VIII.

scholarly journals Protein Measurement of Particulate and Solubilized Ovine Liver Membranes

1992 ◽  
Vol 29 (6) ◽  
pp. 659-662 ◽  
Author(s):  
Anastassia Hatzoglou ◽  
John Prekezes ◽  
Marie Tsami ◽  
Elias Castanas
Keyword(s):  
Protein Concentration ◽  
Cell Surface Receptor ◽  
Blue Dye ◽  
Cellular Membranes ◽  
Surface Receptor ◽  
Liver Membranes ◽  
Biuret Method ◽  
Concentration Measurements ◽  
Triton X

Protein concentration measurements are critical in biochemical work with cellular membranes, including the determination of cell surface receptor concentration in human malignant tissues obtained at surgery or after biopsy. In this study we compared the results of protein concentration measurements in ovine liver cellular membranes using either particulate preparations or membranes solubilized with four different detergents. In all cases protein was determined by two different indirect methods (Lowry's Folin phenol method and Bradford's Coomassie Brilliant Blue dye binding method) and compared to the direct biuret method. Our results indicate that the direct biuret method gives the highest protein concentrations followed by the method of Lowry. Maximal concentrations (approaching those obtained by the direct biuret method) were obtained after membrane solubilization with Triton X-100 (3–5%). It is suggested that either the direct biuret method (whenever protein concentrations permit it) or the method of Lowry after solubilization of membranes with Triton X-100 (3–5%) should be used preferentially for the determination of membrane protein samples.

scholarly journals Determination of the concentration of low molecular fraction of Сandida Albicans proteins by ELISA method at subcutaneous introduction in candidiasis therapy

2020 ◽  
Vol 11 (SPL4) ◽  
pp. 2896-2899
Author(s):  
Mykola Rybalkin ◽  
Tatiana Diadiun ◽  
Natalia Khokhlenkova ◽  
Serhiy Stepanenko ◽  
Nataliia Dvinskykh
Keyword(s):  
Molecular Weight ◽  
Therapeutic Effect ◽  
Protein Concentration ◽  
Subcutaneous Administration ◽  
Weight Fraction ◽  
Low Molecular Weight ◽  
Elisa Method ◽  
Defence Mechanisms ◽  
Molecular Fraction

The purpose of this work was to determine the C. albicans protein concentration at the subcutaneous introduction. The therapeutic  effect of C. albicans proteins has been studied in white mice.  Animals were injected  intraperitoneally with a suspension of C. albicans fungus.  After five days and repeatedly, after 14 days, mice were infected subcutaneously  with proteins of Candida cells of volume 0.2 mL. Fourteen days after each injection, the determination of the protective functions of the animal body has been carried out by the titer of specific C. albicans antibodies. According to the data obtained during studies on the treatment of candidiasis, it was found that in the subcutaneous administration after the first and second injection with a concentration of low molecular weight fraction of the C. albicans protein of 1, 2, 3, 4, and 5 mg/mL antibodies titers have doubled, indicating a lack of activation of the body's protective functions. Proteins of low molecular weight fraction of C. albicans with concentrations 1, 2, 3, 4, and 5 mg/mL do not activate the body's defence mechanisms.

scholarly journals Evaluation of urinary proteins in healty full-term neonates by SDS-PAGE

2005 ◽  
Vol 51 ◽  
pp. 9-14
Author(s):  
Snezhana Palchevska ◽  
Velibor Tasik ◽  
Petar Korneti ◽  
Georgi Shestakov ◽  
Svetlana Tsekovska
Keyword(s):  
Molecular Weight ◽  
Full Term ◽  
Urine Samples ◽  
Low Molecular Weight ◽  
Urinary Proteins ◽  
Term Neonates ◽  
Molecular Weights ◽  
Electrophoretic Profile ◽  
Sds Page

The pattern of urinary proteins in healthy full-term neonates was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), coupled with determination of few parameters related to urinary protein excretion. Twenty healthy full-term neonates were included in the investigation. Five urine samples from each subject were collected on days 3, 7, 14, 21 and 28 after birth. Determination of total proteins was performed using turbidimetric method with sulfosalicylic acid. Urinary creatinine concentration was determined by Jaffe method. Urinary proteins were separated by horizontal gradient SDS-PAGE according to Görg. The highest values for total urinary proteins and for protein/creatinine ratio were detected in urine samples excreted on days 3 and 7 after birth. Three types of SDS-PAG electrophoretic profiles were observed. The first type of electrophoretic profile was characterized by the presence of proteins of mixed glomerular and tubular origin with molecular weights from 10 to 160 kDa. Typical for the second type of electrophoretic profile was the presence of two faint fractions with molecular weights of 78 and 90 kDa and several intensive low molecular weight fractions (14-67 kDa). In the third type of electrophoretic profile only low molecular weight proteins (10-67 kDa) were detected in all five urine samples. These findings express the transitory immaturity of the glomerular filter and tubular protein reabsorbing system of the newborn kidney. Apparently, the tubular protein handling normalizes later than the glomerular filtration of proteins.

Improved SDS-PAGE Molecular Weight Determination of a Succinylated Limed Ossein Gelatin

2002 ◽  
Vol 7 (3) ◽  
pp. 195-209 ◽  
Author(s):  
M. Kaur ◽  
A. Hardman ◽  
C. D. Melia ◽  
K. Jumel ◽  
S. Higginbottom
Keyword(s):  
Molecular Weight ◽  
Molecular Weight Determination ◽  
Sds Page

Fractionation of Proteins in Surimi Waste Water Using Membrane Filtration

2012 ◽  
Author(s):  
Raweewan Tacharatanamanee ◽  
Kitti Cherdrungsi ◽  
Wirote Youravong
Keyword(s):  
Waste Water ◽  
Molecular Weight ◽  
Pore Size ◽  
Membrane Filtration ◽  
Narrow Range ◽  
Protein Recovery ◽  
Membrane Selectivity ◽  
Sds Page ◽  
Key Factor

The study was conducted to determine the feasibility of using ultrafiltration and microfiltration for fractionation of proteins in surimi waste water. The results from this study showed that the molecular weight of the soluble proteins in surimi waste water was in the range of 10–120 kDa. Ultrafiltration surimi waste water with using membrane with MWCO 100 and 300 kDa could not fractionate these proteins since most of the proteins were retained in retentate. This result suggested that fouling formed during ultrafiltration played an important role in determination of the membrane selectivity. Improving the fouling problem may be the key factor enhancing membrane selectivity. Fractionation of proteins from this waste by using microfiltration with membranes pore size 0.22, 0.45, and 1 μm was also studied. Although some proteins could penetrate through the membrane, the results from SDS–PAGE showed that the protein profiles in the retentate and permeates did not differ, indicating that these membranes also could not be used for fractionation. This may be due to the large pore size of the membranes and the narrow range of the molecular weight of these proteins. Key words: Surimi waste water, protein recovery, fractionation, ultrafiltration, microfiltration

scholarly journals PURIFICATION AND MOLECULAR WEIGHT DETERMINATION OF KERATINASE ISOLATED FROM STREPTOMYCES MALAYSIENSIS

2017 ◽  
Vol 9 (10) ◽  
pp. 154
Author(s):  
M. Pavani ◽  
G. Girijasankar ◽  
K. Mallika ◽  
G. Vidhya Sagar
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sole Source ◽  
Molecular Weight Determination ◽  
Termite Mound ◽  
Phylogenetic Tree Analysis ◽  
Sds Page ◽  
Streptomyces Malaysiensis

Objective: The aim of the present study was to purify and determine the molecular weight of keratinase isolated from Streptomyces malaysiensis.Methods: For that purpose purification was done using ammonium sulphate and Sephadex-LH 100 column chromatography. Further, the fractions were pooled and subjected to molecular weight determination using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE).Results: The obtained results showed keratinase with 47.57% recovery, 3.5-fold purification and an estimated molecular mass of 27,000 Da. Keratinase showed an optimal activity at 60 οC and pH 8. Keratinase activity of the purified product was assayed with feather powder as a substrate. The isolated strain was identified as Streptomyces malaysiensis based on phylogenetic tree analysis. The strain isolated from termite mound soil showed the highest keratinase activity, which could be considered a microorganism of environmental origin.Conclusion: The production of keratinase on simple media with feathers as sole source allowing its production from the cheap substrate and a commercial production with low production cost. Stability in the presence of detergents, surfactants and solvents make this keratinase extremely useful for a biotechnological process involving keratin.

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